Structure-activity mapping of ARHGAP36 reveals regulatory roles for its GAP homology and C-terminal domains.

ARHGAP36 is an atypical Rho GTPase-activating protein (GAP) family member that drives both spinal cord development and tumorigenesis, acting in part through an N-terminal motif that suppresses protein kinase A and activates Gli transcription factors. ARHGAP36 also contains isoform-specific N-termina...

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Autores principales:	Patricia R Nano, Taylor K Johnson, Takamasa Kudo, Nancie A Mooney, Jun Ni, Janos Demeter, Peter K Jackson, James K Chen
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/26c7e95d7e2c4d55b0b5a8130ee17cf1
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spelling	oai:doaj.org-article:26c7e95d7e2c4d55b0b5a8130ee17cf12021-11-25T06:19:10ZStructure-activity mapping of ARHGAP36 reveals regulatory roles for its GAP homology and C-terminal domains.1932-620310.1371/journal.pone.0251684https://doaj.org/article/26c7e95d7e2c4d55b0b5a8130ee17cf12021-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0251684https://doaj.org/toc/1932-6203ARHGAP36 is an atypical Rho GTPase-activating protein (GAP) family member that drives both spinal cord development and tumorigenesis, acting in part through an N-terminal motif that suppresses protein kinase A and activates Gli transcription factors. ARHGAP36 also contains isoform-specific N-terminal sequences, a central GAP-like module, and a unique C-terminal domain, and the functions of these regions remain unknown. Here we have mapped the ARHGAP36 structure-activity landscape using a deep sequencing-based mutagenesis screen and truncation mutant analyses. Using this approach, we have discovered several residues in the GAP homology domain that are essential for Gli activation and a role for the C-terminal domain in counteracting an N-terminal autoinhibitory motif that is present in certain ARHGAP36 isoforms. In addition, each of these sites modulates ARHGAP36 recruitment to the plasma membrane or primary cilium. Through comparative proteomics, we also have identified proteins that preferentially interact with active ARHGAP36, and we demonstrate that one binding partner, prolyl oligopeptidase-like protein, is a novel ARHGAP36 antagonist. Our work reveals multiple modes of ARHGAP36 regulation and establishes an experimental framework that can be applied towards other signaling proteins.Patricia R NanoTaylor K JohnsonTakamasa KudoNancie A MooneyJun NiJanos DemeterPeter K JacksonJames K ChenPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 5, p e0251684 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Patricia R Nano Taylor K Johnson Takamasa Kudo Nancie A Mooney Jun Ni Janos Demeter Peter K Jackson James K Chen Structure-activity mapping of ARHGAP36 reveals regulatory roles for its GAP homology and C-terminal domains.
description	ARHGAP36 is an atypical Rho GTPase-activating protein (GAP) family member that drives both spinal cord development and tumorigenesis, acting in part through an N-terminal motif that suppresses protein kinase A and activates Gli transcription factors. ARHGAP36 also contains isoform-specific N-terminal sequences, a central GAP-like module, and a unique C-terminal domain, and the functions of these regions remain unknown. Here we have mapped the ARHGAP36 structure-activity landscape using a deep sequencing-based mutagenesis screen and truncation mutant analyses. Using this approach, we have discovered several residues in the GAP homology domain that are essential for Gli activation and a role for the C-terminal domain in counteracting an N-terminal autoinhibitory motif that is present in certain ARHGAP36 isoforms. In addition, each of these sites modulates ARHGAP36 recruitment to the plasma membrane or primary cilium. Through comparative proteomics, we also have identified proteins that preferentially interact with active ARHGAP36, and we demonstrate that one binding partner, prolyl oligopeptidase-like protein, is a novel ARHGAP36 antagonist. Our work reveals multiple modes of ARHGAP36 regulation and establishes an experimental framework that can be applied towards other signaling proteins.
format	article
author	Patricia R Nano Taylor K Johnson Takamasa Kudo Nancie A Mooney Jun Ni Janos Demeter Peter K Jackson James K Chen
author_facet	Patricia R Nano Taylor K Johnson Takamasa Kudo Nancie A Mooney Jun Ni Janos Demeter Peter K Jackson James K Chen
author_sort	Patricia R Nano
title	Structure-activity mapping of ARHGAP36 reveals regulatory roles for its GAP homology and C-terminal domains.
title_short	Structure-activity mapping of ARHGAP36 reveals regulatory roles for its GAP homology and C-terminal domains.
title_full	Structure-activity mapping of ARHGAP36 reveals regulatory roles for its GAP homology and C-terminal domains.
title_fullStr	Structure-activity mapping of ARHGAP36 reveals regulatory roles for its GAP homology and C-terminal domains.
title_full_unstemmed	Structure-activity mapping of ARHGAP36 reveals regulatory roles for its GAP homology and C-terminal domains.
title_sort	structure-activity mapping of arhgap36 reveals regulatory roles for its gap homology and c-terminal domains.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/26c7e95d7e2c4d55b0b5a8130ee17cf1
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Structure-activity mapping of ARHGAP36 reveals regulatory roles for its GAP homology and C-terminal domains.

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